Pharmaceutical compositions for promoting the growth of gram-positive bacilli and increasing the acidity in the vagina and the use thereof

ABSTRACT

The present invention relates to a pharmaceutical formulation for stimulating the growth of gram-positive bacilli and increasing the acidity in the vagina which comprises sucrose and/to maltose, to the use of certain sucrose and/or maltose, in preparing the pharmaceutical formulation for stimulating the growth of gram-positive bacilli and increasing the acidity in the vagina, in particular to a method of stimulating the growth of gram-positive bacilli and increasing the acidity in the vagina, treating the reduction of gram-positive bacilli and the lowness of acidity in vagina as well as the vaginitis and the disturbance of vaginal bacterioflora accompanying the reduction of gram-positive bacilli, especially bacterial vaginal disease.

This application is a continuation-in-part application of Ser. No.09/577,858, now U.S. Pat. No. 6,632,796 which is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

This invention relates to pharmaceutical compositions containingsaccharides as active ingredients for promoting the growth ofGram-positive bacilli and increasing the acidity in the vagina, to theuse of particular saccharides in the preparation of compositions forpromoting the growth of Gram-positive bacilli and increasing the acidityin the vagina, and especially to a method of promoting the growth ofGram-positive bacilli and increasing the acidity in the vagina, treatingdecreased levels of Gram-positive bacilli and decreased levels ofacidity in the vagina, treating vaginitis and disturbances of thevaginal bacterial flora accompanying the reduction of Gram-positivebacilli, especially bacterial vaginosis.

BACKGROUND OF THE INVENTION

High acidity in female vagina is one important anti-infective mechanismof the vagina and is of great significance for vaginal health.Lactobacilli and other Gram-positive bacilli that can produce and resistacids serve an important role in maintaining the normal acidity in thevagina by keeping the vaginal pH value in the range from 4.0 to 4.6.They are the physiological bacterial flora of vagina, whereasGram-negative bacilli, Gram-negative cocci, and Gram-positive cocci arerelatively less abundant in the healthy vagina.

When the Gram-positive bacilli are reduced or disappear in vagina,vaginal pH value rises and disturbance of vaginal bacterial floraresults from abnormal increases of Gram-negative bacilli, Gram-positivecocci and Gram-negative cocci, which can cause harm to the human bodyand lead to a range of diseases. The most typical condition resultingfrom altered vaginal flora is bacterial vaginosis (BV). BV ischaracterized by the reduction or even disappearance of Lactobacillusand other Gram-positive bacilli in the vagina, accompanied by decreasedacidity (pH value>4.6) in the vagina, and abnormal increases of suchbacteria as Gram-negative bacilli including Gardnerella, Bacteroides andmotile-curved bacilli; Gram-negative cocci such as Veillonella; andGram-positive cocci such as Streptococcus. Such changes in the bacterialflora can cause vaginal secretions to exhibit an unpleasant odor, andmay be associated with pruritus of vulva, and symptoms. In addition, BVmay also be related to IUGR [1], PTL, PROM [2], abortion, and obstetricinfections such as chorio-amnionitis, puerperal endometritis, vaginalwall phlegmon after hysterectomy, female upper genital tract infection(salpingitis), and urinary infection, etc. [3]. A high rate of morbidityis associated with vaginal bacterial flora disturbance. According to onereport, about 45% or more vaginitis cases result from disturbance ofvaginal bacterial flora [3], and 4-15% of American female students inuniversities suffer from bacterial vaginosis [4], which has led toserious compromise to health and quality of life.

There are few options for treatment of reduced Gram-positive bacillicolonization, decreased vaginal acidity, and related disturbances ofvaginal bacterial flora, vaginitis, and bacterial vaginosis. Therapeuticoptions currently include:

1) Antibacterial drugs which are used to suppress the growth ofGram-negative bacilli and other abnormal bacteria. These most commonlyinclude clindamycin and metronidazole [5-6]. These drugs suppress thebacteria that are abnormally increased in the vagina but may also affectthe Gram-positive bacilli. After administration of these drugs, theGram-positive bacilli (lactobacilli) can not be restored very well, andit is very difficult to lower the pH value in the vagina to normallevel.

2) Lactic acid-containing pharmaceutical compositions. Vaginalsecretions from patients suffering from bacterial vaginosis haveelevated pH values. Swedish researchers used lactic acid gel for theimprovement and recovery of the low-acidity conditions in vagina, andreported that this treatment can restore the Gram-positive bacilli(lactobacilli) in the vaginas of some of the patients [7]. But the studyalso showed that the lactic acid pharmaceutical preparation is lesseffective than the antibacterial drugs [8].

3) Lactobacillus preparations. Most of the Gram-positive bacilli invagina are lactobacillus. If there is disturbance of vaginal bacterialflora, the lactobacilli will be reduced or disappear, and Gram-negativebacilli, Gram-negative, and Gram-positive cocci, will increase. TheGram-positive bacilli in the vagina of some patients can be restored bydirectly adding lactobacilli in the vagina [9]. However, stablecolonization is generally not achieved. Moreover, it is difficult tomaintain viability of the lactobacillus preparations during storage,with viable counts in such preparations decreasing during storage,compromising their useful shelf life [10].

The international Patent Application WO94/02148 discloses apharmaceutical composition for treating vulvitis and vulvovaginitis, andindicates that such compositions can promote restoration of vaginalepithelium tissues while alleviating the symptoms. Its preferredcomposition comprises 7 to 8 active substances. Some preferredcompositions may contain 3.0-15.0% (by weight/volume) lactose orglucose. As mentioned in page 5 lines 8-10 of the publishedspecification, the lactose or glucose contained in these compositions isused as carbon source. But this application does not mention thatsaccharides can be used solely as the effective component for treatingvulvitis and vulvovaginitis, and nor does it disclose explicitly orimplicitly that the disclosed compositions can stimulate the growth ofGram-positive bacilli in vagina. Furthermore, it does not indicate thatany other kinds of sugar can be used as active components of acomposition for treating related vaginal diseases. Besides, as mentionedin page 5 lines 17-18 of the specification, this application emphasizesthat it is important for the pH value of the compositions be between 2and 3.5.

The U.S. Pat. No. 3,860,707 teaches a method for treating trichomonalvaginitis and monilial vaginitis. This method comprises administeringlactulose into the vagina. This patent also indicates that lactulose canbe administered after being mixed with some carriers such as glucose,lactose and galactose, wherein lactulose is required to have aconcentration as high as 50%, and the mixture also contains 5% lactose,8% galactose as carriers, as mentioned in column 1 lines 51-55 andcolumn 5 lines 1-5 of the patent specification. The quantity oflactulose is 4-10 grams administered with each dose, which is taken onceor twice daily, as shown in Column 4 Lines 63-66 of the specification.But this patent does not describe the treatment effectiveness onbacterial vaginosis or other vaginal diseases different from monilialvaginitis, nor does it suggest that the lactulose of low or mediumconcentration (2.5-17%) and small dosage (daily total amount 0.24-2.1grams) would be able to stimulate the growth of Gram-positive bacillusand increase the acidity of the vagina. Furthermore, it fails toindicate whether any saccharide other than lactulose has treatmenteffects.

European Patent Application EP-A-0257007 discloses a pharmaceuticalcomposition containing lactic acid and buffering substances andsubstrate to support growth of lactobacillus, which can be used toimprove micro-environment in vagina and suppress the growth of harmfulbacteria in the vagina, so as to facilitate the growth of lactobacilli.This patent application discloses that glycogen or lactose can be usedas the said substrate. But as mentioned in Column 6 Line 10-14 of thespecification, the main ingredient of this composition is lactic acid.The lactic acid and the glycogen and/or lactose are incorporated in aratio by weight of from 20:1 down to 500:1, and the content of glycogenand/or lactose is only 0.1-0.166% (W/V). It also stresses that the pHvalue of the pharmaceutical composition should be adjusted to 3.5 to4.0, which is very important. The in vitro experimental resultsdisclosed in this application show that this composition can effectivelyand selectively kill pathogenic bacteria, and lactobacilli can survivein this composition for a longer time than the pathogenicmicroorganisms. But no test in vitro or vivo shows that this compositioncan stimulate the growth of lactobacilli or produce acids. Nor does thisapplication mention the treatment effect of glycogen or lactose or anyother saccharides when they are used separately as active ingredients.

GB21 12285A discloses a lotion composition for cleaning the vagina. Itis a buffering liquid comprising acetic acid or lactic acid plus sodiumacetate with a pH value of 5.71 to 6.2 as shown in the examples. It alsocontains nutrients to support the growth of lactobacillus [1-2% (W/V)glucose and unsaturated fatty acid]. It also generally mentionsinclusion of mono- and/or disaccharides. The main therapeutic mechanismof this composition is that the buffering lotion comprising acetic acidor lactic acid plus sodium acetate can selectively suppress pathogensand not suppress lactobacilli. As shown in claims 2 and 3 and the invitro test data of this application, this lotion can effectivelysuppress many kinds of pathogens, and lactobacilli survive in thiscomposition for a longer time. No data from test in vitro or vivoindicates that this lotion has an activity of promoting the growth oflactobacilli and producing acid, nor does it indicate the treatmenteffect of glucose or any other sugar when used separately as activeingredients. This application teaches that lactobacilli regulate pHvalue in vagina to about 5.8, as shown in Page 1 Lines 20-23 of thespecification, which is strongly contradicted by most knowledgeableinvestigators.

The above-mentioned pharmaceutical compositions disclosed in patentapplications EP-A-0257007 and GB2112285A which contain lactic acid,acetic acid and other selective inhibitors as main active ingredientshave strong suppressive action on pathogens but no explicit suppressiveaction on lactobacilli, although they may indirectly facilitate thegrowth of vaginal lactobacilli. These compositions themselves, however,cannot directly promote significant growth of lactobacilli, and onlyregulate vaginal acidity for a short time. Therefore, it remains verydifficult to restore the physiological conditions dominated by theGram-positive bacillus-flora and to restore the vaginal acidity to itsnormal value.

The object of the present invention is to provide a composition forpromoting the growth of Gram-positive bacilli and the production ofacids, and thus increase the acidity in the vagina. Another object ofthe present invention is to provide a method by using such compositionfor reversing the reduction of Gram-positive bacilli, lack of vaginalacidity and treating related vaginal diseases.

In order to seek a composition which is effective in promoting thegrowth of Gram-positive bacilli, producing acid, and enhancing theacidity in the vagina, the inventor has conducted an extensive study,performed tests by using various pharmaceutical compositions known inthe prior art, and has not found any compositions promoting the growthof Gram-positive bacilli among the existing compositions. After repeatedtests and intensive study, the inventor has found very surprisingly thatsucrose and maltose both have a strong effect in promoting the growth ofGram-positive bacilli and producing acids if they are presented in aconcentration and at a pH value in specific ranges. Combined in vitroculturing experiments show that the two saccharides can stimulate thegrowth of Gram-positive bacilli, increasing their numbers significantly.To our surprise, although pH values above 4.6 in vagina are consideredun-physiologic, and most of state-of-the-art technologies stress thatthe pharmaceutical compositions used in vagina must have a pH valueequal to 4.0 or below, the inventor has discovered that, if they have pHvalue between 4.1 and 7.2, and especially above pH 5.0, sucrose andmaltose can stimulate the growth of Gram-positive bacilli of womenvagina and the production of acid, and are able to decrease the pH valuein vagina to less than 4.6. However, if they have pH values of 4.0 orless, they do not exert significant growth-promoting effects onGram-positive bacilli nor upon acid production and the pH of the vaginacan rarely be reduced to below 4.6. Based on the above discoveries andfurther study, the inventor has completed the present invention.

SUMMARY OF THE INVENTION

The present invention provides a water-based pharmaceutical compositionfor promoting the growth of Gram-positive bacilli and increasing theacidity in the vagina comprising, based on the volume of thecomposition, 2.5% to 17% (W/V) of sucrose and/or maltose and optionallyone or more saccharides selected from the group consisted of glucose,fructose, galactose, mannose, lactose, lactulose, mycose, cellobiose,melibiose, melitose, malto-oligosaccharide, iso-malto-oligosaccharideand oligo-fructose, dextrim, starch and glycogen, at pH-value of 4.1-7.2adjusted with pharmaceutically acceptable acid or alkali, optionally apharmaceutically acceptable viscous base, and optionally an effectiveamount of anti-fungal and/or an anti-bacterial agents.

The invention also provides a use of sucrose and/or maltose as activeingredient in the preparation of pharmaceutical compositions forpromoting the growth of Gram-positive bacilli and increasing the acidityin the vagina.

The invention also relates to a method for promoting the growth ofGram-positive bacilli and increasing the acidity in the vagina,comprising administering to the subject in need of such treatment atherapeutically effective amount of the pharmaceutical compositionaccording to the present invention.

The present application further relates to a composition for treating apatient suffering from vaginitis, a disturbance of the vaginal bacterialflora or bacterial vaginosis, wherein said vaginitis, disturbance of thevaginal bacterial flora or bacterial vaginosis are accompanied with areduction of the number of Gram-positive bacilli, said compositioncomprising:

-   -   (a) Sucrose and/or maltose in a concentration of from about 2.5%        to about 17% w/v based on the total volume of the composition,    -   (b) An anti-fungal drug in a concentration of from about 0.0001%        to about 5% w/v based on the total volume of the composition,        and    -   (c) A sufficient amount of a pharmaceutically acceptable acid or        alkali, which results in a pH of the composition from about 4.1        to about 7.2.

The above-mentioned pharmaceutical composition, use and method oftreatment according to the invention are useful for the reversal of thereduced numbers of vaginal Gram-positive bacilli, decreased vaginalacidity, as well as for treating vaginitis and the disturbance ofvaginal bacterial flora accompanied with the reduction in numbers ofGram-positive bacilli, especially bacterial vaginosis.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, this invention relates to water-based pharmaceuticalcompositions with pH values between 4.1 and 7.2 intended to promote thegrowth of Gram-positive bacilli and enhance vaginal acidity, and whichcontains 2.5-17% (WIV) of one or more of such saccharides as definedabove.

The compositions according to the present invention may contain one or amixture of two or several of such saccharides as defined above. Anyhexose used in the invention is of D-type. Starch used in the inventionmay be amylose or amylopectin. The preferred saccharides are thosehaving low price and abundant sources.

According to this invention, the pharmaceutical composition contains atotal content of 2.5-17% (W/V) of saccharides, especially 2.5-16% (W/V),preferably 8-14% (W/V), more preferably 10-13% (W/V), and mostpreferably 10-12% (W/V). However, the content of the disaccharides otherthan sucrose and maltose mixed with sucrose and/or maltose must notexceed [(17—content of sucrose and/or maltose) % (W/V)]. The maximumcontent of hexoses mixed with sucrose and/or maltose should be less than[0.42×(17—disaccharides content) % (W/V)].

The weight/volume content (W/V) mentioned in the context of thisapplication refers to the grams of the specified component in 100milliliters of the composition.

The compositions formulated according to the preferred contents ofsaccharides are useful for treating the patients with any vaginalillness states, especially with severe illness (with pH value of vaginalsecretion greater than 5.0, and when the vaginal Gram smear proves thatthere are few or no Gram-positive bacilli). The compositions with acontent of saccharides below 8% (W/V) are applicable to the patientssuffering from mild diseases (with a vaginal pH value of greater than4.6, and the vaginal Gram smear proves that there are Gram-positivebacilli, but in which the Gram positive bacilli are fewer in number thanthe Gram-negative bacilli, Gram-negative cocci, or Gram-positive cocci).

The pH value of the pharmaceutical compositions of this invention isbetween 4.1 and 7.2, with the optimum pH value between 4.5 and 6.5. ThepH value of these compositions can be adjusted by adding anypharmaceutically acceptable acid or alkali, of which the preferredchoices are acetic acid, lactic acid, or sodium hydroxide. The natureand concentration of such acid or alkali can be readily determined by aperson skilled in the art.

According to this invention, the composition may contain a viscous base.One example of such base is Xanthan Gum with concentration of 1.0-2.2%(W/V), and preferably of 1.4-2.0% (W/V). Xanthan Gum is able to keep thesugar in uniform contact with vaginal mucosa and retain the productwithin the vaginal vault for a long time due to its high adherence andstability against changes of temperature and pH values, thus permit thecompositions to promote the growth of Gram-positive bacilli andincreasing the production of acid in vagina.

The composition can also be formulated into hydro-gel form or ointmentwith other suitable viscous carrier bases, auxiliaries well known to aperson skilled in the art.

According to this invention, the composition also may not containviscous base, it may be administered by means of intravaginal tamponsaturated with the liquid composition. The intravaginal tampon may becomposed of cotton ball, gauze ball, ribbon gauze, etc. In thisembodiment, the composition according to the present invention can alsostimulate the growth of Gram-positive bacilli and increase the acidityin the vagina. The preferred use of the composition of this inventiondoes requires that the composition of the invention stay in the vaginafor some time before it can stimulate the growth of Gram-positivebacilli and produce vaginal acidity in the vagina. Therefore as a lotionwithout a viscous base, the composition can not exhibit its therapeuticeffect very well.

The saccharide(s) is/are the essential basic active components of thecomposition of the invention, and can fulfill the object of thisinvention when used with suitable pharmaceutically acceptable carriers.But these saccharides can produce a better treatment effect if it iscombined with minor amount of amino-acid, vitamin or other similarsubstances, or yeast extract rich in amino-acids and vitamins. Thevaginal secretion naturally contains sufficient amino-acids andvitamins. Such amino-acids or vitamins are not available in conventionalin vitro experiments and should be added when in vitro experiments arecarried out for the composition of the invention.

Sucrose and/or maltose promote the growth of vaginal Gram-positivebacilli and increase vaginal acidity, which would inhibit abnormalvaginal bacterial flora except Gram-positive bacilli. In this way thereduced vaginal Gram-positive bacilli and the disturbance of vaginalflora, vaginitis, or bacterial vaginosis could be cured.

However, the inventor noticed further that in seldom cases Candidacoexisted with abnormal vaginal bacterial flora or even BV flora.Although increased vaginal Gram-positive bacilli and vaginal acidity didinhibit abnormal vaginal bacterial flora, it didn't inhibit Candida.Candida might grow better in vagina as a result of inhibition ofabnormal vaginal bacterial flora. In order to treat such cases ofreduced vaginal Gram-positive bacilli or disturbance of vaginal flora orvaginits or bacterial vaginosis, the inventor conducted further studiesand this new composition comprising sucrose and/or maltose andanti-fungi drugs was invented. As the anti-fungi drugs may also beeffective against bacteria, the preferred content of anti-fungi drug inthis invented composition is in low concentration so to avoid inhibitingGram-positive bacilli. Compared with some anti-fungi creams available inmarket whose anti-fungi drugs concentration may be as high as 100 to1000 times the drug's MIC against Candidal strains, in this inventedcomposition the content of anti-fungi drug may be as low as 0.1˜10 timesthe drug's MIC.

The present application further relates to a composition for treating apatient suffering from vaginitis, a disturbance of the vaginal bacterialflora or bacterial vaginosis, wherein said vaginitis, disturbance of thevaginal bacterial flora or bacterial vaginosis are accompanied with areduction of the number of Gram-positive bacilli, said compositioncomprising:

-   -   (a) Sucrose and/or maltose in a concentration of from about 2.5%        to about 17% w/v based on the total volume of the composition,    -   (b) Anti-fungi drug in a concentration of from about 0.0001% to        about 5% w/v based on the total volume of the composition, and    -   (c) A sufficient amount of a pharmaceutically acceptable acid or        alkali, which results in a pH of the composition from about 4.1        to about 7.2.

The above mentioned composition may further comprise one or moresaccharides from the group consisting of glucose, fructose, galactose,mannose, lactose, lactulose, mycose, cellobiose, melibiose, melitose,malto-oligosaccaride, iso-malto-oligosaccharide and oligo-fructose,dextrin, starch and glycogen.

Preferably, the content of sucrose and/or maltose in said composition isfrom about 8% to about 14% w/v, the content of anti-fungi drug in saidcomposition is from about 0.001% to about 0.5% w/v. More preferably, thecontent of anti-fungi drug in said composition is in the range of 0.1 to10 times of the drug's MIC against Candidal strains.

The anti-fungi drug in the above mentioned composition is selected fromthe following group: Fluconazole, Terconazole, Tioconazole,Butoconazole, Ketoconazole, Itraconazole, Econazole, Miconazole, andCannitracin. Preferably, the antifungi drug is selected fromFluconazole, Terconazole, and Tioconazole.

Said composition may be in the form of hydro-gel or ointment, or in theform of liquid for preparing intravaginal tampon. Preferably, thecomposition is in the form of hydro-gel or ointment, said compositionfurther comprises pharmaceutically acceptable viscous base, preferably,said pharmaceutically acceptable viscous base is Xanthan gum.

The present application also relates to a method for treating a patientsuffering from vaginitis, a disturbance of the vaginal bacterial floraor bacterial vaginosis, wherein said vaginitis, disturbance of thevaginal bacterial flora or bacterial vaginosis are accompanied with areduction of the number of Gram-positive bacilli, said method comprisingvaginally administering to a subject in need of such treatment atherapeutically effective amount of a composition comprising:

-   -   (a) Sucrose and/or maltose in a concentration of from about 2.5%        to about 17% w/v based on the total volume of the composition,    -   (b)Anti-fungi drug in a concentration of from about 0.0001% to        about 5% w/v based on the total volume of the composition, and    -   (c) A sufficient amount of a pharmaceutically acceptable acid or        alkali, which results in a pH of the composition from about 4.1        to about 7.2;    -   wherein said administration promotes selective growth of        gram-positive bacilli in the vagina of said subject.

The present application further relates to a method for treating apatient suffering from vaginitis, a disturbance of the vaginal bacterialflora or bacterial vaginosis, wherein said vaginitis, disturbance of thevaginal bacterial flora or bacterial vaginosis are accompanied with areduction of the number of Gram-positive bacilli, said methodcomprising, simultaneously or sequentially, vaginally administering to asubject in need of such treatment a therapeutically effective amount ofa composition (A) and composition (B),

-   -   Composition (A) comprising:        -   a) Sucrose and/or maltose in a concentration of from about            2.5% to about 17% w/v based on the total volume of the            composition, and        -   b) A sufficient amount of a pharmaceutically acceptable acid            or alkali, which results in a pH of the composition from            about 4.1 to about 7.2;    -   Composition (B) comprising:        -   (a) Anti-fungi drug in a concentration of from about 0.0001%            to about 5% w/v based on the total volume of the            composition, and        -   (b) A sufficient amount of a pharmaceutically acceptable            acid or alkali, which results in a pH of the composition            from about 4.1 to about 7.2.

Preferably, composition (A) and composition (B) are vaginallyadministered to a subject in need of such treatment simultaneously atherapeutically effective amount of a composition (A) and composition(B).

The composition of the invention may also contain one or moreanti-bacterial agents that can suppress or kill Gram-negative bacteriabut exert no effect or only exert slight effect on Gram-positivebacilli. In this embodiment, the composition of the invention may havean increased efficacy for treating vaginal infection or inflammation.Such anti-bacterial agents may include, but are not limited topolymyxin, metronidazole or aztreonam.

The composition of the invention can be prepared according to theprocesses known to those skilled in the art.

If the saccharides used include little or no starch, such saccharideshould be mixed with viscous auxiliary substances homogeneously, andthen distilled water is added into the mixture, which are then stirredto dissolve the saccharide and swell the viscous auxiliary substancesuntil a homogeneous viscous gel is formed. If starch is included, it issufficient to heat directly the mixture of saccharides and water to forma paste. In the latter case, viscous auxiliary substances may be addedor not. For adjusting the pH to a predetermined value, lactic acid orsodium hydroxide solution is added prior to sterilization treatment.Alternatively, sterilization treatment is performed first, followed byadjustment of pH. For sterilization, intermittent sterilization may beused, with the detailed steps described as follows: sterilizing at 80°C. for 30 minutes, keeping at 36° C. for 8-12 hours, sterilizing at 80°C. for 30 minutes, keeping at 36° C. for 8-12 hours and finallysterilizing at 80° C. for 30 minutes. Alternatively, a high-pressuresterilization may be performed for 15-20 minutes at 116° C. A solutionof saccharides may also be sterilized by filtering the solution. Thenthe sterilized saccharide solution may be added to a viscous base in theform of hydro-gel that has been sterilized at high pressure.

The composition of the invention also may be made into a solution bydissolving the saccharides in water. The solution can be administered toa patient by means of an intravaginal tampon soaked in it.

The composition of this invention uses sugar substances as activeingredients, and has good stability during storage, but preferably it isstored under refrigeration or in cool place. The near neutral pH valueof the composition is also helpful in stabilizing the sugar components.

The present invention also relates to the use of one or more of suchsaccharides as defined above as active ingredients in the preparation ofa medicament for promoting the growth of Gram-positive bacilli andincreasing the acidity in the vagina. This invention uses D-type hexose,either amylose or amylopectin.

The medicament prepared according to the use of the invention can beused for promoting the growth of Gram-positive bacilli and increasingthe acidity in the vagina, and reversing decreased numbers ofGram-positive bacilli, diminished vaginal acidity (pH value above 4.6)as well as treating vaginitis and the disturbance of vaginal bacterialflora accompanied with the reduction of Gram-positive bacilli,especially bacterial vaginosis.

The experiments in vitro and in vivo have proven that the composition ofthe invention can strongly stimulate the growth of Gram-positive bacilliand increase the acidity in the vagina, and can be used for the reversalof the reduction of Gram-positive bacilli, diminished vaginal acidityand treatment of vaginitis and the disturbance of vaginal bacterialflora accompanied with reduction of Gram-positive bacilli, especiallybacterial vaginosis.

Therefore, this invention also relates to a method for promoting thegrowth of Gram-positive bacilli and increasing vaginal acidity,reversing the reduction of Gram-positive bacilli, increasing vaginalacidity and treatment of vaginitis and the disturbance of vaginalbacterial flora that accompanied with reduction of Gram-positivebacilli, especially bacterial vaginosis, wherein the subject in need ofsuch treatment is given a medically-effective amount of thepharmaceutical composition according to the invention.

The administration method of the composition is to administer thecomposition locally inside the vagina. The composition of the inventioncontaining a tissue viscous base or other carriers may be applieddirectly to the lumen of the vagina. If the composition of the inventionis in the form of a solution, an intravaginal tampon is soaked in thesolution, then the tampon is placed inside the vagina.

For the composition and method of treatment according to the invention,the medicament is administered according to the following dosage. Forthe composition of this invention containing 8-14% (W/V) of saccharidesas active ingredients, such composition is applied inside the vagina 1-3times daily in doses of 1-5 ml, with the total sugar amount controlledto 0.08-2.1 grams daily dosage, generally applied before sleep at nightor after arising in the morning, with an additional dose applied at noonfor a few patients. For the patients suffering from severely abnormalvaginal bacterial flora and with the pH value of the vaginal secretiongreater than 5.0, and if the vaginal Gram smear shows few or noGram-positive bacilli, more extensive treatment is required with thesugar amount above 0.8 grams daily. For the patients having less severedisease with the pH value of vaginal secretion between 5.0 and 4.6, orif the vaginal Gram smear reveals Gram-positive bacilli, but in lesserabundance than that of any of Gram-negative bacilli, Gram-negativecocci, or Gram-positive cocci, a smaller dosage is used and the totalsugar amount is limited to 0.8 grams or less.

During treatment with this composition, clinical symptoms may beobserved daily and the vaginal pH value checked for change. Moreover,the vaginal Gram smear may be performed in order to check the change ofbacterial flora and adjust the treatment accordingly if necessary.Generally, the composition of this invention can produce remarkabletherapeutic effects 1-3 days after beginning of use, with symptomsimproved significantly even disappearing, and pH values in the vaginareduced to normal levels and the Gram-positive bacilli in the vaginarestored to dominance in the vaginal bacterial flora, at which timetherapy with the composition should be stopped or the dosage be reduced,or the treatment continued at a low maintenance dosage.

For the method of this invention, the patients are provided with thecomposition containing only the saccharides of this invention as activeingredients, or the composition containing the saccharides, anti-fungalagent and/or anti-bacterial agent. Alternatively, the compositioncontaining the saccharides of this invention as its active ingredientsis administered in conjunction with suitable anti-fungal agent oranti-bacterial agent. For the latter case, the composition of thisinvention can be administered simultaneously with the anti-fungal and/oranti-bacterial agent or before/after the administration of theanti-fungal and/or anti-bacterial agent.

After the administration of this composition, the clinical symptoms ofpatients can be alleviated quickly, the numbers of Gram-positive bacilliare increased in the vagina, vaginal acidity is raised with the pH valuereduced to 4.0-4.6, while the Gram-negative bacilli, Gram-negative cocciand other harmful abnormal bacteria are reduced substantially or evendisappear. The composition of this invention is easy to prepare and touse with reliable effects.

EXPERIMENTAL EXAMPLE 1

Experiment in vitro with the composition of this invention: The effectof the composition in promoting the growth of Gram-positive bacilli andthe production of acids.

Method:

-   (1) The preparation of the composition: Sucrose was used to prepare    the following composition according to the method mentioned above:    sucrose 10.0% (W/V), yeast extract 1.0% (W/V), Xanthan gum 1.6%    (W/V), pH 5.0; then filled the composition into the tubes after    sterilization, with each tube containing 5 ml, and pre-reduced for    use.-   (2) Specimen suspension: Vaginal secretion was taken from one of the    patients suffering from typical bacterial vaginosis with a cotton    swab, then the swab was washed in 2 ml sterilized Trypcase-soy Broth    immediately, and thus the specimen suspension was ready. The vaginal    Gram smear showed few Gram-positive bacilli but an abundance of    Gram-negative bacilli, negative cocci and positive cocci.-   (3) The above-mentioned specimen suspension was inoculated    immediately into the tubes containing the above-mentioned    composition, 10 microliter for each tube, mixed homogeneously. The    tubes were placed in an incubator for cultivation, at 37° C.,    anaerobically. Then, culture samples were taken from the tubes at 10    hours and 24 hours later respectively. The Gram smears of the    samples were observed and the pH values of the samples were    measured.    Results:

As shown in Table 1, although the Gram smear of the specimen showed fewGram-positive bacilli, the Gram positive bacilli grew remarkably in thecomposition of this invention after specimen suspension inoculated andcultivated. Meanwhile the pH value of the composition decreased.

TABLE 1 The Effect of the Composition of this Invention in Promoting theGrowth of Gram-positive Bacilli and Producing Acids Sacchride Bacteria10 hours- 24 hours- contained specimen in pH of culture culture incomposition suspension the composition Bacteria pH Bacteria pH SucroseG + b*, −** 5.0 G + b, +++ 5.0 G + b, ++++ 4.0 G − b, ++++ G − b, ++ G −b, ++ G − c, +++ G + c, + G − c, + G + C, ++ G + C, ++ G + C, ++ *G + b:Gram-positive bacilli, G − b: Gram-negative bacilli; G + c:Gram-positive cocci, G − c: Gram-negative cocci. **− No or less than onebacterium per field of version under oil-immersion lens; +: 1-9 bacteriaper field of vision under oil-immersion lens; ++: About 10-99 bacteriaper field of vision under oil-immersion lens; +++: About 100 bacteria ormore per field vision under oil-immersion lens, even uncountable; ++++:Bacteria clumped or aggregated.Conclusion:

The composition of this invention has the effect of promoting the growthof Gram-positive bacilli and the production of the acids.

EXPERIMENT EXAMPLE 2

Experiment in vitro with the compositions of this invention: thecomparison of the effects of compositions containing sucrose or maltose,and those containing other saccharides in promoting the growth ofGram-positive bacilli and the production of acids

Method:

-   (1)The preparation of the compositions: Different saccharides were    used respectively to prepare the following compositions according to    the methods mentioned above: glucose, fructose, galactose, mannose,    maltose, sucrose, lactose, lactulose, mycose, cellobiose, melibiose,    melitose, malto-oligosaccharide, isomaltooligosaccharide and    fructooligosaccharide, dextrin, starch, and glycogen:    -   A. 5% glucose, 1.0% yeast extract, 1.6% xanthan gum, pH adjusted        to 6.2;    -   B. 5% fructose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   C. 5% galactose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   D. 5% mannose, 1.0% yeast extract, 1.6% xanthan gum, pH adjusted        to 6.2;    -   E. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   F. 10.0% sucrose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   G. 10.0% lactose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   H. 10.0% lactulose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   I. 10.0% cellobiose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   J. 10.0% mycose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   K. 10.0% melibiose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   L. 10.0% melitose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   M. 10.0% maltooligosaccharide, 1.0% yeast extract, 1.6% xanthan        gum, pH adjusted to 6.2;    -   N. 10.0% isomaltooligosaccharide, 1.0% yeast extract, 1.6%        xanthan gum, pH adjusted to 6.2;    -   O. 10.0% fructooligosaccharide, 1.0% yeast extract, 1.6% xanthan        gum, pH adjusted to 6.2;    -   P. 10.0% dextrin, 1.0% yeast extract, 0.5% xanthan gum, pH        adjusted to 6.2;    -   Q. 10.0% starch, 1.0% yeast extract, pH adjusted to 6.2;    -   R. 10.0% glycogen, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2.-   (2)The preparation of the test tubes: The compositions prepared    above were filled into test tubes, with each tube containing 5 ml,    sterilized, and kept until use for experiment.-   (3) Specimen suspension: Vaginal secretion was taken from one of the    patients suffering from typical bacterial vagmosis with a cotton    swab, then the swab was washed in 2 ml sterilized Trypcase-soy Broth    immediately, and thus the specimen suspension was ready. The vaginal    Gram smear showed few Gram-positive bacilli but an abundance of    Gram-negative bacilli, negative cocci and positive cocci.-   (4)The specimen suspension was inoculated immediately into the tubes    containing the above-mentioned compositions, 10 microliter for each    tube, mixed homogeneously. Then the tubes were placed in a candle    jar and cultivated at 37° C. After 24 hours and 48 hours' culture,    culture samples were taken respectively from each of the tubes, then    the Gram smears of the samples were observed and the pH values of    culture samples were tested.    Results:-   (1)As shown in Table 2, although there was few Gram positive bacilli    in the vaginal secretion specimen, the Gram positive bacilli grew    remarkably in the compositions containing different sugars of this    invention after the compositions were inoculated with specimen    suspension and cultivated for 24 hours or 48 hours. Meanwhile the pH    values in most of the composition tubes decreased to different    levels. These results indicate that sucrose, maltose, or any other    kind of saccharides compatible to them according to this invention    exerts respectively effect in promoting the growth of Gram-positive    bacilli.-   (2)As shown in Table 2, the fact that the pH values in 10% maltose    tube and in 10% sucrose tube were lower than that of the other tubes    after 24 or 48 hours culture indicates that maltose and sucrose are    metabolized rapidly by bacteria. And maltose and sucrose exert    better effect in promoting the growth of Gram-positive bacilli and    the production of acids than that of the other kinds of    oligosaccharides.

TABLE 2 Effects of Sucrose, Maltose and Other Saccharides onGram-positive Bacilli Growth and Acids-producing Sacchride Bacteria in24 hours- 48 hours- contained in specimen PH of the culture culturecomposition suspension composition Bacteria pH Bacteria pH Glucose G +b, − 6.2 G + b, ++ 6.4 G + b, +++ 6.2 G − b, ++++ G − b, + G − b, + G −c, +++ G − c, − G − c, − G + c, ++ G + c, − C + c, ++ Fructose G + b, −6.2 G + b, ++ 5.4-5.8 G + b, +++ 5.4-5.8 G − b, ++++ G − b, + G − b, + G− c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, + Galactose G + b,− 6.2 G + b, ++ 6.7 G + b, + 6.7 G − b, ++++ G − b, − G − b, + G − c,+++ G − c, − G − c, − G + c, ++ G + c, ++ G + c, ++ Mannose G + b, − 6.2G + b, ++ 6.4 G + b, +++ 5.8 G − b, ++++ G − b, + G − b, + G − c, +++ G− c, − G − c, − G + c, ++ G + c, ++ G + c, + Maltose G + b, − 6.2 G + b,++ 4.6-48 G + b, +++ 4.4-4.6 G − b, ++++ G − b, +++ G − b, ++ G − c, +++G − c, − G − c, − G + c, ++ G + c, ++ G + c, + Sucrose G + b, − 6.2 G +b, ++ 4.8-5.1 G + b, +++ 4.6-4.8 G − b, ++++ G − b, + G − b, ++ G − c,+++ G − c, − G − c, − G + c, ++ G + c, + G + c, + Lactose G + b, − 6.2G + b, ++ 5.8 G + b, ++ 5.1-5.4 G − b, ++++ G − b, + G − b, + G − c, +++G − c, − G − c, − G + c, ++ G + c, +++ G + c, + Lactulose G + b, − 6.2G + b, +++ 5.8-6.2 G + b, ++ 6.2 G − b, ++++ G − b, + G − b, + G − c,+++ G − c, − G − c, − G + c, ++ G + c, + G + c, + Cellobiose G + b, −6.2 G + b, ++ 5.8 G + b, ++ 5.8 G − b, ++++ G − b, + G − b, + G − c, +++G − c, − G − c, − G + c, ++ G + c, + G + c, + Mycose G + b, − 6.2 G + b,++ 4.8 G + b, +++ 4.6-4.8 G − b, ++++ G − b, + G − b, ++ G − c, +++ G −c, − G − c, − G + c, ++ G + c, + G + c, + Melibiose G + b, − 6.2 G +b, + 6.2 G + b, ++ 5.8 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G −c, + G − c, − G + c, ++ G + c, + G + c, + Melitose G + b, − 6.2 G + b,++ 6.7 G + b, +++ 5.8 G − b, ++++ G − b, + G − b, ++ G − c, +++ G − c, +G − c, − G + c, ++ G + c, + G + c, + Maltooligo- G + b, − 6.2 G + b, ++5.8 G + b, +++ 5.8 Sacchride G − b, ++++ G − b, + G − b, + G − c, +++ G− c, − G − c, − G + c, ++ G + c, ++ G + c, ++ Fructooligo- G + b, − 6.2G + b, +++ 5.8 G + b, +++ 6.2 saccharide G − b, ++++ G − b, ++ G − b, +G − c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, − Isomaltooligo-G + b, − 6.2 G + b, ++ 6.2 G + b, +++ 5.8 Saccharide G − b, ++++ G − b,++ G − b, ++ G − c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, +Dextrin G + b, − 6.2 G + b, +++ 6.2 G + b, ++ 5.8 G − b, ++++ G − b, ++G − b, + G − c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, − StarchG + b, − 6.2 G + b, ++ 6.7 G + b, ++ 6.2 G − b, ++++ G − b, ++ G − b, ++G − c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, + Glycogen G + b,− 6.2 G + b, ++ 6.4 G + b, +++ 6.2 G − b, ++++ G − b, ++ G − b, ++ G −c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, +Conclusion:

The sugar components contained in the compositions of this invention canbe sucrose, maltose or the both or mixtures of sucrose and/or maltoseand other saccharides as defined above.

EXPERIMENT EXAMPLE 3

Experiment in vitro with the compositions of this invention: The effectsof the compositions of this invention in promoting the growth ofGram-positive bacilli and the production of acids.

Methods:

-   (1) The preparation of the compositions: Maltose and sucrose were    used respectively to prepare the following composition according to    the methods mentioned above:    -   A. 2.5% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   B. 2.5% sucrose, 1.0 yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;-   (2) The preparation of the test tubes: Compositions as prepared    above were filled into the test tubes, with each tube containing 5    ml, sterilized, and kept until use for experiment.-   (3) Specimen suspension: Vaginal secretion was taken from one of the    patients suffering from typical bacterial vaginosis with a cotton    swab, then the swab was washed in 2 ml sterilized Trypcase-soy Broth    immediately, and thus the specimen suspension was ready. The vaginal    Gram smear showed few Gram-positive bacilli but an abundance of    Gram-negative bacilli, negative cocci and positive cocci.-   (4) The above-mentioned specimen suspension was inoculated    immediately into the tubes containing the above-mentioned    compositions, 10 microliter for each tube, mixed homogeneously. Then    the tubes were placed in a candle jar and cultivated at 37° C. After    24 hours and 48 hours' culture, culture samples were taken    respectively from each of the tubes, then the Gram smears of the    samples were observed and the pH values of culture samples were    tested.    Results:

As shown in Table 3, although there was few Gram positive bacilli in thevaginal secretion specimen, the Gram positive bacilli grew in thecompositions containing different sugars of this invention after thecompositions were inoculated with specimen suspension and cultivated for24 hours or 48 hours. It indicates that 2.5% of sucrose and 2.5% ofmaltose have the effects in promoting the growth of Gram-positivebacilli.

TABLE 3 The Effect of the Compositions of This Invention in Promotingthe Growth of Gram-positive Bacilli and the Production of AcidsSaccharde 24 hours- 48 hours- contained in Bacteria in PH of the cultureculture composition specimen suspension composition Bacteria pH BacteriapH Maltose G + b, − 6.2 G + b, + 7.0 G + b, +++ 6.4 G − b, ++++ G − b,+++ G − b, + G − c, +++ G − c, − G − c, − G + c, ++ G + c, − G + c, +Sucrose G + b, − 6.2 G + b, + 6.2 G + b, + 6.7 G − b, ++++ G − b, + G −b, ++ G − c, +++ G − c, − G − c, − G + c, ++ G + c, + G + c, +Conclusion:

The compositions of this invention containing 2.5% sucrose or maltoseexert certain effects in promoting the growth of Gram-positive bacilli.

EXPERIMENT EXAMPLE 4

Experiment in vitro with the compositions of this invention: effects ofthe compositions of this invention in promoting the growth ofGram-positive bacilli and the production of acids.

Method:

-   (1)The preparation of the compositions: Maltose and sucrose were    used respectively to prepare the following composition in the    methods mentioned above:    -   A. 17.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;    -   B. 17.0% sucrose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.2;-   (2) The preparation of the test tubes: Compositions prepared above    were filled into the test tubes, with each tube containing 5 ml,    sterilized, and kept until use for experiment.-   (3) Specimen suspension: Vaginal secretion was taken from one of the    patients suffering from typical bacterial vaginosis with a cotton    swab, then the swab was washed in 2 ml sterilized Trypcase-soy Broth    immediately, and thus the specimen suspension was ready. The vaginal    Gram smear showed few Gram-positive bacilli but an abundance of    Gram-negative bacilli, negative cocci and positive cocci.-   (4) The above-mentioned specimen suspension was inoculated    immediately into the tubes containing the above-prepared    compositions, 10 microliter for each tube, mixed homogeneously. Then    the tubes were placed in a candle jar and cultivated at 37° C. After    24 hours and 48 hours' culture, culture samples were taken    respectively from each of the tubes, then the Gram smears of the    samples were observed and the pH values of culture samples were    tested.    Results:

As shown in Table 4, although there was few Gram positive bacilli in thevaginal secretion specimen, numerous Gram positive bacilli grew in thecompositions of this invention after the compositions were inoculatedwith specimen suspension and cultivated for 24 hours or 48 hours.Meanwhile the pH values of the composition tubes decreased remarkably.These results indicate that compositions containing 17% of sugars ofthis invention exert remarkable effect in promoting the growth ofGram-positive bacilli.

TABLE 4 Selective Promoting Effect on the Growth of Gram-positiveBacilli and the Production of acids Saccharide 24 hours 48 hourscontained in the Bacteria in pH of the culture culture preparationspecimen inoculated preparation Bacteria pH Bacteria pH Maltose G + b, −6.2 G + b, +++ 5.4 G + b, ++ 5.1 G − b, ++++ G − b, + G − b, ++ G − c,+++ G − c, − G − c, − G + c, ++ G + c, + G + c, ++ Sucrose G + b, − 6.2G + b, ++ 6.2 G + b, ++ 5.4 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G− c, − G − c, − G + c, ++ G + c, + G + c, ++Conclusion:

The compositions of this invention containing 17% sucrose or maltoseexert effects in promoting the growth of Gram-positive bacilli.

EXPERIMENT EXAMPLE 5

Experiment in vitro with the compositions of this invention: Theinfluence of different pH values on the effects of the compositions ofthis invention on the growth of Gram-positive bacilli and the productionof acids.

Method:

-   (1)The preparation of the compositions: Maltose was used to prepare    the following compositions according to the method mentioned above:    -   A. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 4.4;    -   B. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 4.8;    -   C. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 5.1;    -   D. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 5.4;    -   E. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 5.8;    -   F. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.4;    -   G. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 6.8;    -   H. 10.0% maltose, 1.0% yeast extract, 1.6% xanthan gum, pH        adjusted to 7.2;-   (2)The preparation of the test tubes: Compositions prepared above    were filled into the test tubes, with each tube containing 5 ml,    sterilized, and kept until use for experiments.-   (3)Specimen suspension: Vaginal secretion was taken from one of the    patients suffering from typical bacterial vaginosis with a cotton    swab, then the swab was washed in 2 ml sterilized Trypcase-soy Broth    immediately, and thus the specimen suspension was ready. The vaginal    Gram smear showed few Gram-positive bacilli but an abundance of    Gram-negative bacilli, negative cocci and positive cocci.-   (4)The above-mentioned specimen suspension was inoculated    immediately into the tubes containing the above-mentioned    compositions, 10 microliter for each tube, mixed homogeneously. Then    the tubes were placed in a candle jar and cultivated at 37° C. After    24 hours and 48 hours' culture, culture samples were taken    respectively from each of the tubes, then the Gram smears of the    samples were observed and the pH values of culture samples were    tested.    Results:

As shown in Table 5,

-   (1) When the pH value of the composition of this invention was 4.4,    a few of Gram-positive bacilli grew in the composition after 48    hours' culture.-   (2) When the pH values of the compositions of this invention were    5.4-7.2, the Gram-positive bacilli grew very well. Gram-positive    bacilli grew and the pH values decreased after 24 hours' culture and    the bacilli were numerous and the pH values decreases to about    4.1-4.4 after 48 hours' culture.

TABLE 5 The Influence of Different pH values of the Compositions of ThisInvention on the Treatment Effects Saccharide Bacteria in 24 hours 48hours contained in the specimen pH of the culture culture compositioninoculated composition Bacteria pH Bacteria pH Maltose G + b, − 4.4 G +b, −˜+ 4.4 G + b, −˜+ 4.4 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G −c, − G − c, − G + c, ++ G + c, − G + c, − Maltose G + b, − 4.8 G + b, +4.8 G + b, + 4.4-4.8 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G − c, −G − c, − G + c, ++ G + c, + G + c, + Maltose G + b, − 5.1 G + b, + 4.6G + b, ++ 4.4-4.6 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G − c, − G− c, − G + c, ++ G + c, + G + c, ++ Maltose G + b, − 5.4 G + b, +++ 5.4G + b, ++ 4.1-4.4 G − b, ++++ G − b, + G − b, ++ G − c, +++ G − c, − G −c, − G + c, ++ G + c, + G + c, − Maltose G + b, − 5.8 G + b, ++ 4.8 G +b, +++ 4.1 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G − c, − G − c, −G + c, ++ G + c, + G + c, + Maltose G + b, − 6.4 G + b, + 4.4 G + b, +++4.1 G − b, ++++ G − b, ++ G − b, ++ G − c, +++ G − c, − G − c, − G + c,++ G + c, + G + c, + Maltose G + b, − 6.8 G + b, ++ 5.4 G + b, +++ 4.4 G− b, ++++ G − b, ++ G − b, ++ G − c, +++ G − c, − G − c, − G + c, ++ G +c, − G + c, + Maltose G + b, − 7.2 G + b, ++ 5.4 G + b, +++ 4.1 G − b,++++ G − b, + G − b, ++ G − c, +++ G − c, − G − c, − G + c, ++ G + c, −G + c, −Conclusion:

In this in vitro experiment, the compositions of this invention with pHvalues of 5.4-7.2 exerted stronger effects in promoting the growth ofGram-positive bacilli.

EXPERIMENT EXAMPLE 6

Case Report

Ms. Jiang, female, aged 30. Her vaginal secretions had exhibitedunpleasant fish-odor accompanying pruritus of vulvae for 2 years.Examined and diagnosed as “bacterial vaginosis” by the present inventor.The patient was treated with a composition containing 12% (W/V) oflactose and having pH value of 5.0. The drug was intravaginallyadministered, 3 ml for each time, once a day. After the treatmentcontinued successively for three days, the pH value of the vaginalsecretions of this patient decreased from 5.4 to 4.6. But the Gramsmears showed that the majority of the bacteria were still Gram-negativebacilli, and there were also a lot of positive cocci and positivebacilli. Thus, the drug for treatment was changed to a composition ofthis invention containing 12% (W/V) of maltose, with the pH value 5.0.10 hours after the administration of 3 ml of the latter composition, thepH value of the vaginal secretions of the patient decreased to 3.8. Thenthe dose was reduced to 2 ml of the composition for each time, once aday. The pH values of the vaginal swabs were maintained at normal levels(pH<4.6). Afterwards, the illness of this patient relapsed after themenstruation several times, and the abnormal bacterial flora and thedecreased vaginal acidity of this patient after the menstruationrestored to normal states by the intravaginal administration of thecomposition of this invention containing 12% (W/V) of maltose.

EXPERIMENT EXAMPLE 7

Experiment in vivo with the composition of the present invention: theinfluences of the composition of this invention on the vaginal bacterialflora of rhesus macaques.

Experimental Method:

-   (1) Animals: Took the vaginal swabs of ten female macaques, tested    the pH values and carried out Gram smears examination. The results    showed that in eight rhesus macaques, the pH values of the vaginal    swabs were greater than 4.6 and Gram smears revealed few    Gram-positive bacilli, a lot of Gram-negative bacilli and negative    cocci, alike the bacterial flora of bacterial vaginosis in human    beings. In the two left rhesus macaques, the pH values of the    vaginal swabs were lower than 4.6 and the bacteria revealed on Gram    smears were Gram-positive bacilli, alike the bacterial flora of the    normal vaginal bacterial flora of human beings.-   (2) In the 8 rhesus macaques, the vaginal bacterial flora were the    type of bacterial vaginosis of human beings. 3 of them were excluded    from the experiment because of menstruation. The other 5 rhesus    macaques were included in the experiment.-   (3) Composition: A composition of this invention was used, which    contained 12% (W/V) of sucrose, 1% (W/V) of yeast extract, 1.6%    (W/V) of xanthan gum and its' pH value was 5.1.-   (4) Administration: 1 ml of the above-mentioned composition of this    invention was administrated intravaginally, twice a day. When most    of the vaginal bacteria was changed to Gram-positive bacilli, the    dose was reduced or the treatment stopped. In this experiment, 1    macaque received the composition only once, 2 macaques twice, and    the other 2 macaques three times.-   (5) Result examination: Took the vaginal swabs before each dosage    was administered to detect the pH values and to perform Gram smears    examination. Thus the change of the vaginal bacterial flora of the    macaques was monitored.    Result:

After the treatment with the composition of the present invention, thevaginal pH values were reduced to 3.8-4.1 and the vaginal bacterialfloras were changed to be dominated with Gram-positive bacilli in allthe five macaques. This result indicates that the composition of thisinvention has very strong effects in vivo in promoting the growth ofGram-positive bacilli and the production of acids.

ADVANTAGES OF THIS INVENTION COMPARED TO THE PRIOR ART

Advantages Compared to Anti-bacterial Treatments:

Anti-bacterial drugs, based on the views of etiology, control theabnormal bacteria that grow excessively and cause pathological reactionby killing or suppressing such bacteria. This method has the followingshortcomings: (1) After repeated treatments with anti-bacterial drugs,the bacteria may gain drug-resistances which may lead to the failure ofantibacterial therapy. (2) Anti-bacterial treatment may result in thesuperinfection by drug-resistant bacteria. (3) Anti-bacterial drugs maybe allergic to human body and may have other kinds of adverse effects toskin or vaginal mucous membrane. Compositions containing lactic acid oracetic acid or other selective bacterial inhibitors as its activeingredients, for example, the compositions described in the patentapplications Nos. GB21 12285A and EP-A-0257007, exert strong prohibitioneffects on the pathogens and have no remarkable prohibition effects onlactobacilli. Thus they may exert indirectly the favorable effects onthe growth of lactobacilli in the vagina. The compositions themselves,however, cannot directly and remarkably promote the growth oflactobacilli in the vagina. The effects of these compositions inincreasing the acidity in the vagina last only for a short period oftime, and it is very hard for the physiological Gram-positive bacilli torestore and dominate over the vaginal bacterial flora.

The compositions of this invention can stimulate the growth ofGram-positive bacilli and increase acidity in the vagina, and thussuppress Gram-negative bacilli, Gram negative cocci, and Gram-positivecocci thereby. Seen from above analysis, the technologies andcompositions of this invention actually enhance the naturalphysiological anti-disease mechanisms in the vagina and fundamentallyavoid and overcome the disadvantages of the disturbance of vaginalbacterial flora by anti-bacterial treatment, therefore have remarkableadvantages.

Advantages Compared to Lactobacilli Preparations:

Firstly, the effects of lactobacilli preparations in the treatment ofsevere disturbance of the vaginal bacterial flora, such as typicalbacterial vaginosis. are often variable. Secondly, the lactobacillipreparations are more difficult to produce and store in addition to itshigh production cost. The compositions of this invention have a lowercost of production and longer effective period, and are significantlysuperior over the lactobacilli preparations. Moreover, the compositionsof this invention improve the condition of the local micro-habitat inthe vagina thus promote the growth of the endogenous Gram—positivebacilli in the vagina. It should be better than the direct supplement ofthe exogenous lactobacilli strains.

COMPOSITION EXAMPLES EXAMPLE 1

2.5 g of sucrose and 2.0 g xanthan gum were mixed homogeneously. Then tothe resultant mixture, some distilled water is added while stirring inorder to dissolve the sugar component and swell the xanthan gum tohomogeneous viscous gum. By adding a suitable amount of lactic acid, thepH of the solution was adjusted to 5.0. Distilled water was added untilthe total volume of the solution is equal to 100 ml. The resultingsolution is sterilized by means of intermittent sterilization.

EXAMPLE 2

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1, exceptthat the pH value is adjusted with 0.5 N sodium hydroxide.

Sucrose 17.0% (W/V) Yeast extract powder 1.0% (W/V) Xanthan gum 1.5%(W/V) Distilled water q.s. pH 7.2

EXAMPLE 3

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Sucrose 12.0% (W/V) Yeast extract powder 1.0% (W/V) Xanthan gum 1.0%(W/V) Distilled water q.s. pH 4.1

EXAMPLE 4

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Sucrose 4.0% (W/V) Maltose 5.0% (W/V) Fructose 2.0 (W/V) Histidine 100ppm Methionine 50.0 ppm Riboflavin 0.2 ppm Thiamine 0.2 ppm Nicotinicacid 0.2 ppm Calcium pantothenate 0.2 ppm Xanthan gum 1.5% (W/V)Distilled water q.s. pH 6.4

EXAMPLE 5

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 2.5% (W/V) Xanthan gum 1.6% (W/V) Distilled water q.s. pH 6.8

EXAMPLE 6

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 17.0% (W/V) Xanthan gum 1.6% (W/V) Yeast extract 1.0% (W/V)Distilled water q.s. pH 5.4

EXAMPLE 7

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 13.0% (W/V) Yeast extract 1.0% (W/V) Xanthan gum 1.6% (W/V)Distilled water q.s. pH 6.4

EXAMPLE 8

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 10.0% (W/V) Glucose 2.0% (W/V) Yeast extract 1.0% (W/V) Xanthangum 1.6% (W/V) Distilled water q.s. pH 4.8

EXAMPLE 9

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Sucrose 12.0% (W/V) Glucose 1.0% (W/V) Yeast extract 0.S% (W/V) Xanthangum 1.7% (W/V) Distilled water q.s. pH 5.6

EXAMPLE 10

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Sucrose 10.0% (W/V) Glucose 1.0% (W/V) Histidine 100 ppm Methionine 50.0ppm Riboflavin 0.2 ppm Thiamine 0.2 ppm Nicotinic acid 0.2 ppm Calciumpantothenate 0.2 ppm Xanthan gum 1.8% (W/V) Distilled water q.s. pH 6.5

EXAMPLE 11

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 10.0% (W/V) Ketoconazole 2.0% (W/V) Yeast extract 1.0% (W/V)Xanthan gum 1.5% (W/V) Distilled water q.s. PH 6.0

EXAMPLE 12

100 ml of the composition in the following formulation was preparedsubstantially according to the method as described in Example 1.

Maltose 9.0% (W/V) Polymyxin E 0.5% (W/V) Yeast extract 1.0% (W/V)Xanthan gum 2.2% (W/V) Distilled water q.s. pH 6.3

FORMULATION EXAMPLE 1

100 ml of the composition in the following formulation is preparedbasically in the methods as described in Example 1.

Sucrose 9.0% (W/V) Fluconazole 0.01% (W/V) Xanthan gum 1.5% (W/V)Distilled water 100 ml PH 7.0

FORMULATION EXAMPLE 2

100 ml of the composition in the following formulation is preparedbasically in the methods as described in Example 1.

Sucrose 9.0% (W/V) Terconazole 0.04% (W/V) Xanthan gum 2.2% (W/V)Distilled water 100 ml PH 7.0

The compositions of this invention are simple in components and can beeasily manufactured at low cost.

EXPERIMENTAL EXAMPLE 1

Materials and Methods

-   -   1. Materials:        -   Sucrose Broth: 10.0% sucrose, 2.0% yeast extract,            K₂HPO₄0.5%, MgSO₄0.05%, MnSO₄0.1%, pH adjusted to 6.2,            sterilized.        -   Anti-fungi drug: Fluconazole        -   Clinically isolated Lactobacillus strains: Lactobacillus            acidophilus, Lactobacillus crispatus, and Lactobacillus            jensenii. Three lactobacillus suspensions were prepared            separately from each of these three lactobacillus strains.        -   Clinically isolated Candida strains: Candida albicans and            Candida tropicalis. Two Candidal suspensions were prepared            separately from each of these two candida strains.    -   2. Methods:        -   Grouping: 24 tubes were divided into 3 lactobacillus strain            groups and 1 Candidal group, as indicated in Table 1.

TABLE 1 Four Groups of Different Microbial Strains and FluconazoleConcentrations Concentration of Fluconazole (ug/ml) Microbes of eachgroup Tube1 Tube2 Tube3 Tube4 Tube5 Control-tube Group 1 400 200 100 5025 0 L. acidophilus C. albicans C. tropicalis Group 2 400 200 100 50 250 L. crispatus C. albicans C. tropicalis Group 3 400 200 100 50 25 0 L.jensenii C. albicans C. tropicalis Group 4 400 200 100 50 25 0 C.albicans C. tropicalis

-   -   -   There were six tubes in each group and each tube contained            2ml Sucrose Broth and different concentration of            Fluconazole, as shown in Table 1.        -   The turbidity values of three lactobacillus suspensions and            two Candidal suspensions were tested by Turbidimeter and            adjusted to 0.3. 50 ul of each prepared lactobacillus            suspension and Candidal suspension were separately added            into tubes according to Table 1.        -   All of these experimental tubes were put into candle jars as            soon as the lactobacillus suspensions and candidal            suspensions were added, and then incubated at 37° C. for 18            hours.        -   After 18 hours' incubation the turbidity of each tube was            examined and Gram-stained smear of cultural fluid of            corresponding tube was observed under microscopy to check            microbes.            Results

The results are shown in Table 2.

-   -   1. The cultural fluids were turbid in three Lactobacillus groups        that indicated microbial growth. Lactobacilli were observed on        the Gram-stained smear of the cultural fluid of these three        experimental groups. Lactobacilli and Candida were observed on        the Gram-stained smear of the cultural fluid of control tubes.        These results showed that 400 ug/ml of Fluconazole didn't        inhibit the growth of three Lactobacillus strains but 25ug/ml or        greater concentration of Fluconazole inhibited the growth of two        Candidal strains.

TABLE 2 The Microbial Growth in Each Experimental Tubes Microbial growthin each tube Tube1 Tube2 Tube3 Tube4 Tube5 Control-tube Group 1 400 200100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++ Microbes Lb Lb Lb Lb Lb Lb & CGroup 2 400 200 100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++ Microbes Lb LbLb Lb Lb Lb & C Group 3 400 200 100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++Microbes Lb Lb Lb Lb Lb Lb & C Group 4 400 200 100 50 25 0 Turbidity +/−+/− +/− +/− +/− ++ Microbes − − − − − C “++”: The cultural fluid washeavily turbid; “+”: The cultural fluid was turbid; “+/−”: The turbidityof the cultural fluid was not affirmative; “Lb”: Lactobacilli; “C”:Candida.

-   -   2. In Group 4, the cultural fluids in experimental tubes was        almost clear and no Candida was found. It was turbid in control        tube and Microscopy examination of the cultural fluid revealed        yeast-like organisms. That showed there was much less Candidal        growth in experimental tubes than in control tube. It indicated        that 25 ug/ml or greater concentration of Fluconazole inhibited        the growth of the two Candidal strains.        Discussion

The results of this experiment proved that certain anti-fungi drug suchas Fluconazole could prevent the growth of Candida but didn't inhibitthe growth of Lactobacilli. Thus it is worthwhile to combine theLactobacilli growth-promoting saccharides of this invention withanti-fungi drugs so as to promote the growth of vaginal lactobacilliwhile preventing the potential growth of Candida in the vagina.

Experimental Example 2

Materials and Methods

-   -   1. Materials:        -   Maltose Broth: 10.0% maltose, 2.0% yeast extract, K₂HPO₄            0.5%, MgSO₄ 0.05%, MnSO₄0.1%, pH adjusted to 6.2,            sterilized.        -   Anti-fungi drug: Terconazole        -   Clinically isolated Lactobacillus strains: Lactobacillus            acidophilus, Lactobacillus crispatus, and Lactobacillus            jensenii. Three lactobacillus suspensions were prepared            separately from each of these three lactobacillus strains.

Clinically isolated Candida strains: Candida albicans and Candidatropicalis. Two Candidal suspensions were prepared separately from eachof these two candida strains.

-   -   2. Methods:        -   Grouping: 24 tubes were divided into 3 lactobacillus strain            groups and 1 Candidal group, as indicated in Table 1.

TABLE 1 Four Groups of Different Microbial Strains and TerconazoleConcentrations Concentration of Terconazole (ug/ml) Microbes in eachgroup Tube1 Tube2 Tube3 Tube4 Tube5 Control-tube Group 1 400 200 100 5025 0 L. acidophilus C. albicans C. tropicalis Group 2 400 200 100 50 250 L. crispatus C. albicans C. tropicalis Group 3 400 200 100 50 25 0 L.jensenii C. albicans C. tropicalis Group 4 400 200 100 50 25 0 C.albicans C. tropicalis

-   -   -   There were six tubes in each group and each tube contained 2            ml Maltose Broth and different concentration of Terconazole,            as shown in Table 1.        -   The turbidity values of three lactobacillus suspensions and            two Candidal suspensions were tested by Turbidimeter and            adjusted to 0.3. 50 ul of each prepared lactobacillus            suspension and Candidal suspension were separately added            into tubes according to Table 1.        -   All of these experimental tubes were put into candle jars as            soon as the lactobacillus suspensions and candidal            suspensions were added, and then incubated at 37° C. for 18            hours.        -   After 18 hours' incubation the turbidity of each tube was            examined and Gram-stained smear of cultural fluid of            corresponding tube was observed under microscopy to check            microbes.            Results

The results are shown in Table 2.

-   -   1. The cultural fluids were turbid in three Lactobacillus groups        that indicated microbial growth. Lactobacilli were observed on        the Gram-stained smear of the cultural fluid of these three        experimental groups. Lactobacilli and Candida were observed on        the Gram-stained smear of the cultural fluid of control tubes.        These results showed that 400 ug/ml of Terconazole didn't        inhibit the growth of three Lactobacillus strains but 25 ug/ml        or greater concentration of Terconazole inhibited the growth of        two Candidal strains.

TABLE 2 The Microbial Growth in Each Experimental Tubes Microbial growthin each tube Tube1 Tube2 Tube3 Tube4 Tube5 Control-tube Group 1 400 200100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++ Microbes Lb Lb Lb Lb Lb Lb & CGroup 2 400 200 100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++ Microbes Lb LbLb Lb Lb Lb & C Group 3 400 200 100 50 25 0 Turbidity ++ ++ ++ ++ ++ ++Microbes Lb Lb Lb Lb Lb Lb & C Group 4 400 200 100 50 25 0 Turbidity +/−+/− +/− +/− +/− ++ Microbes − − − − − C “++”: The cultural fluid washeavily turbid; “+”: The cultural fluid was turbid; “+/−”: The turbidityof the cultural fluid was not affirmative; “Lb”: Lactobacilli; “C”:Candida

-   -   In Group 4, the cultural fluids of experimental tubes seemed        slightly turbid but no organism was found. It was turbid in        control tube and Microscopy examination of the cultural fluid        revealed yeast-like organisms. That showed there was much less        Candidal growth in experimental tubes than in control tube. It        showed that 25 ug/ml or greater concentration of Terconazole        inhibited the growth of the two Candidal strains.        Discussion

The results of this experiment strongly proved that anti-fungi drugssuch as Terconazole could prevent the growth of Candidal strains butdidn't inhibit the growth of Lactobacilli. Thus it is worthwhile tocombine the Lactobacilli growth-promoting saccharides of this inventionwith anti-fungi drugs so as to promote the growth of vaginallactobacilli while preventing the potential growth of Candida in thevagina.

Case Example 3

Ms. Wang, female, aged 38. She had suffered pruritus of vulvae andunpleasant vaginal fish-odor for about 1 year. The inventor checked thepatient's vaginal smear and found that there was mass of Gram-negativebacteria on Gram smears of her vaginal secretions. There were also a fewGram-positive cocci and yeast-like organisms. The vaginal pH value ofthis patient was 5.4. Diagnosed as “bacterial vaginosis” by theinventor, the patient was treated firstly with composition containing10% (W/V) of sucrose. The drug was intra-vaginally administered, 2ml foreach time, once a day. After the treatment continued successively forthree days, the symptoms of the patient relieved significantly and thepH value of the vaginal secretions decreased to 4.0. The Gram smearsshowed a lot of Gram-positive bacilli, a few Gram-negative bacilli, butalso an increased number of yeast-like organisms. Thus the treatment wasstopped. Three days later after stopping the treatment, the patient'svaginal pH value increased to 4.4, Gram-stained vaginal smear showedthere were lots of Gram-negative bacilli again as well as someyeast-like organisms in her vaginal smear. Then the patient was treatedwith the composition of Formulation Example 1 of this invention, 2 mleach time, once a day, for three days. The symptoms of the patient weregone and the pH value of the vaginal secretion was 4.0 after thetreatment. Checking the patient's vaginal flora again and found therewere a lot of Gram-positive bacilli and few Gram-negative bacilli on theGram-stained vaginal smears, no yeast-like organisms were found. Thiscase showed that the combination use of sucrose and Fluconazole couldpromote the growth of Gram-positive-bacilli while preventing the adversegrowth of vaginal yeast-like organisms.

Reference:

-   1) German-M, et al: Genital flora in pregnancy and its association    with intrauterine growth retardation. J-Clin-Microbiol. 1994 Sep:    32(9): 2162-8.-   2) McDonald-HM, et al: Vaginal infection and preterm labour.    Br-J-Obstet-Gynaecol. 1991 May; 98(5): 427-35.-   3) Eschenbach. D A, Clin.Infect.Dis. 1993.1 6(Suppl.4):5282-7.-   4) Thomason J L. Bacterial vaginosis: current review with    indications for asymptomatic therapy. Am J Obstet Gynecol .1991;    165: 1210.-   5) Stein-GE, et al: Placebo-controlled trial of intravaginal    clindamycin 2% cream for the treatment of bacterial. vaginosis.    Ann-Pharmacother. 1993 Nov, 27(11): 1343-5.-   6) Hillier-S L, et al: Efficacy of intravaginal 0.75% metronidazole    gel for the treatment of bacterial vaginosis . Obstet-Gynecol. 1993    Jun; 81(6): 963-7.-   7) Hoster-E, et al: Treatment of bacterial vaginosis in pregnancy    with a lactate gel. Scand-J-Infect-Dis. 1990; 22(5):625-6.-   8) Boeke-A J, et al: Effect of lactic acid suppositories compared    with oral metronidazole and placebo in bacterial vaginosis: a    randomised clinical trial. Genitourrin-Med. 1993 Oct; 69(5): 388-92.-   9) Hallen-A, et al: Treatment of bacterial vaginosis with    lactobacilli. Sex-Transm-Dis. 1992 May-Jun; 19(3):146-8.-   10) Hughes-V L, et al: Microbiologic characteristics of    lactobacillus products used for colonization of the vagina.    Obstet-Gynecol. 1990 Feb; 75(2): 244-8.

1. A method for treating a patient suffering from vaginitis, adisturbance of the vaginal bacterial flora or bacterial vaginosis,wherein said vaginitis, disturbance of the vaginal bacterial flora orbacterial vaginosis are accompanied with a reduction of the number ofGram-positive bacilli, said method comprising vaginally administering toa subject in need of such treatment a therapeutically effective amountof a composition comprising: a) sucrose and/or maltose in aconcentration of from about 2.5% to about 17% w/v based on the totalvolume of the composition, b) anti-fungal drug in a concentration offrom about 0.0001% to about 5% w/v based on the total volume of thecomposition, and c) a sufficient amount of a pharmaceutically acceptableacid or alkali, which results in a pH of the composition from about 4.1to about 7.2; wherein said administration promotes selective growth ofgram-positive bacilli in the vagina of said subject.
 2. A method fortreating a patient suffering from vaginitis, a disturbance of thevaginal bacterial flora or bacterial vaginosis, wherein said vaginitis,disturbance of the vaginal bacterial flora or bacterial vaginosis areaccompanied with a reduction of the number of Gram-positive bacilli,said method comprising, simultaneously or sequentially, vaginallyadministering to a subject in need of such treatment a therapeuticallyeffective amount of a composition (A) and composition (B), whereincomposition (A) comprises: a) sucrose and/or maltose in a concentrationof from about 2.5% to about 17% w/v based on the total volume of thecomposition, and b) a sufficient amount of a pharmaceutically acceptableacid or alkali, which results in a pH of the composition from about 4.1to about 7.2; and composition (B) comprises: (a) anti-fungal drug in aconcentration of from about 0.0001% to about 5% w/v based on the totalvolume of the composition, and (b) a sufficient amount of apharmaceutically acceptable acid or alkali, which results in a pH of thecomposition from about 4.1 to about 7.2.
 3. The method according toclaim 2, wherein composition (A) and composition (B) are vaginallyadministering to a subject in need of such treatment simultaneously. 4.The method according to claim 1, wherein said composition furthercomprising one or more saccharides selected from the group consisting ofglucose, fructose, galactose, mannose, lactose, lactulose, mycose,cellobiose, melibiose, melitose, malto-oligosaccaride,iso-malto-oligosaccharide and oligo-fructose, dextrin, starch andglycogen.
 5. The method according to claim 1, wherein the content ofsucrose and/or maltose in said composition is from about 8% to about 14%w/v.
 6. The method according to claim 1, wherein the content ofanti-fungal drug in said composition is from about 0.001% to about 0.5%w/v.
 7. The method according to claim 1, wherein the anti-fungal drug isselected from the group consisting of Fluconazole, Terconazole,Tioconazole, Butoconazole, Ketoconazole, Itraconazole, Econazole,Miconazole and Cannitracin.
 8. The method according to claim 7, whereinthe anti-fungal drug is selected from the group consisting ofFluconazole, Terconazole and Tioconazole.
 9. The method according toclaim 1, wherein the composition is in the form of hydro-gel orointment, or in the form of liquid for preparing intravaginal tampon.10. The method according to claim 9, wherein the composition is in theform of hydro-gel or ointment and it comprises a pharmaceuticallyacceptable viscous base.
 11. The method according claim 10, wherein thepharmaceutically acceptable viscous base is Xanthan gum.
 12. The methodaccording to claim 2, wherein said composition (A) further comprises oneor more saccharides selected from the group consisting of glucose,fructose, galactose, mannose, lactose, lactulose, mycose, cellobiose,melibiose, melitose, malto-oligosaccaride, iso-malto-oligosaccharide andoligo-fructose, dextrin, starch and glycogen.
 13. The method accordingto claim 2, wherein the content of sucrose and/or maltose in saidcomposition (A) is from about 8% to about 14% w/v.
 14. The methodaccording to claim 2, wherein the content of anti-fungal drug in saidcomposition (B) is from about 0.001% to about 0.5% w/v.
 15. The methodaccording to claim 2, wherein the anti-fungal drug in said composition(B) is selected from the group consisting of Fluconazole, Terconazole,Tioconazole, Butoconazole, Ketoconazole, Itraconazole, Econazole,Miconazole and Cannitracin.
 16. The method according to claim 15,wherein the anti-fungal drug is selected from the group consisting ofFluconazole, Terconazole and Tioconazole.
 17. The method according toclaim 2, wherein the compositions are in the form of hydro-gel orointment, or in the form of liquid for preparing intravaginal tampon.18. The method according to claim 17, wherein the compositions are inthe form of hydro-gel or ointment and it comprises a pharmaceuticallyacceptable viscous base.
 19. The method according to claim 18, whereinthe pharmaceutically acceptable viscous base is Xanthan gum.